WO2017209510A1 - Variant de dunaliella et procédé de production de pigment à l'aide de celui-ci - Google Patents

Variant de dunaliella et procédé de production de pigment à l'aide de celui-ci Download PDF

Info

Publication number
WO2017209510A1
WO2017209510A1 PCT/KR2017/005685 KR2017005685W WO2017209510A1 WO 2017209510 A1 WO2017209510 A1 WO 2017209510A1 KR 2017005685 W KR2017005685 W KR 2017005685W WO 2017209510 A1 WO2017209510 A1 WO 2017209510A1
Authority
WO
WIPO (PCT)
Prior art keywords
algae
dunaliella
pigment
culture
composition
Prior art date
Application number
PCT/KR2017/005685
Other languages
English (en)
Korean (ko)
Inventor
진언선
김민재
Original Assignee
한양대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 한양대학교 산학협력단 filed Critical 한양대학교 산학협력단
Priority to CN201780034310.7A priority Critical patent/CN110225968B/zh
Priority to US16/306,477 priority patent/US11179428B2/en
Publication of WO2017209510A1 publication Critical patent/WO2017209510A1/fr

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/179—Colouring agents, e.g. pigmenting or dyeing agents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
    • A23L17/60—Edible seaweed
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A23L29/035—Organic compounds containing oxygen as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A23L29/045—Organic compounds containing nitrogen as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/40—Colouring or decolouring of foods
    • A23L5/42—Addition of dyes or pigments, e.g. in combination with optical brighteners
    • A23L5/46—Addition of dyes or pigments, e.g. in combination with optical brighteners using dyes or pigments of microbial or algal origin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/047—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates having two or more hydroxy groups, e.g. sorbitol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/02—Algae
    • A61K36/05—Chlorophycota or chlorophyta (green algae), e.g. Chlorella
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • C12N1/125—Unicellular algae isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/202—Algae extracts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/89—Algae ; Processes using algae

Definitions

  • the present invention relates to an alga having a pigment producing ability, a dye composition comprising the alga, and a method for producing the dye.
  • Macular degeneration is a disease that causes visual acuity disorder due to degeneration of the macular tissue, which is located in the center of the inner retina of the eye, and most of the cells are collected in the macula and the image of the object is also the center of the macula. Is in charge of. The most common causes of macular degeneration are age-related macular degeneration and are known to be associated with family history, race, and smoking. Because the macula is responsible for central vision, degeneration occurs in this area, such as decreased vision, central dark spots, and schizophrenia. Macular degeneration is largely divided into non-exudative (dry) and exudative (wet).
  • macular degeneration does not significantly affect visual acuity.
  • the exudative phase which is visible, subretinal hemorrhage, subretinal fluid, or pigmented epithelial detachment, visual acuity develops initially when the location of the lesion is directly under or under the macula.
  • Exudative macular degeneration accounts for about 10-20% of the total macular degeneration, but if left untreated without exudative macular degeneration, visual acuity quickly deteriorates, leading to blindness within two years after diagnosis.
  • Macular pigment is a carotenoid-based oxycarotenoid pigment produced by oxygenation of carotenoids.
  • Xanthophyll examples include lutein or zeaxanthin. Lutein acts as an antioxidant that protects the interior of the eye, which is damaged by the oxygen free radicals that are naturally produced in the body, and kills cancer cells by reducing the growth of blood vessels that supply cancer tumors, breast, colon, and lungs. It is known to have some effects on the prevention of ovarian cancer and skin cancer. Animals cannot produce xanthophyll, and can only obtain it through ingestion of food. These xanthophylls are present with chlorophyll and carotene in the green part of plants, leaves and flowers. Recently, health functional foods for eye health, including xanthophylls, have been in the spotlight.
  • Exemplary marigold flowers are typical sources of zeaxanthin and lutein, and other extracts from higher plants have been studied.
  • the bacteria genetically alter the mechanism of pigment synthesis to produce zeaxanthin and lutein.
  • Research into obtaining these pigments from microalgae has also been conducted.
  • Marigold flower of the conventional raw materials has a disadvantage that it takes a long time to breed the flowers for production, there is a problem that the production cost is high because the production amount is not large compared to the land area for production.
  • the conventional microalgae is a wild type that has not been improved, but the lutein content is constant, but the zeaxanthin content is very low depending on the amount of light.
  • An object of the present invention is to provide a raw material that can replace the xanthophyll used as a raw material of the conventional food or a method that can replace the conventional raw material production method, specifically, xanthophyl production capacity, in particular lutein and zeaxanthin production
  • a composition comprising the same and a method for producing xanthophyll using the same.
  • the present inventors have developed an algae that can solve the insufficient productivity of wild type or existing microalgae by using a variant other than the genetic recombination method which may be a problem in the food industry.
  • the present invention was developed by identifying a mutant strain having a higher yield of macular pigment than a conventional genus of Dunariella, and identifying an optimal pigment production method using the same.
  • the present invention provides an algae of Dunaliella sp. Macular pigment 3 (MP3), KCTC 12990BP.
  • the MP3 algae of the genus Dunariella may have the ability to produce xanthophyll.
  • the MP3 algae of the genus Dunariella are lutein (lutein) and zeaxanthin; And chlorophyll b (chlorophyll b), chlorophyll a (chlorophyll a) and beta-carotene ( ⁇ -carotene) may have the ability to produce one or more pigments selected from the group.
  • the present invention provides a culture of algae ( Dunaliella sp. MP3, KCTC 12990BP) algae genus Dunariella.
  • the present invention provides a pigment composition comprising one or more selected from the group consisting of Dunaliella sp. MP3, KCTC 12990BP algae, a culture of the algae, dried products thereof and extracts thereof. .
  • the present invention also provides Dunalella spp. MP3 ( Dunaliella sp. MP3, KCTC 12990BP) algae, oral administration composition comprising one or more selected from the group consisting of the culture of the algae, dried products and extracts thereof. to provide.
  • Dunalella spp. MP3 Dunaliella sp. MP3, KCTC 12990BP
  • oral administration composition comprising one or more selected from the group consisting of the culture of the algae, dried products and extracts thereof. to provide.
  • the present invention Dunaliella spp. MP3 ( Dunaliella sp. MP3, KCTC 12990BP) algae (algae), the culture of the algae, its dried products and extracts containing one or more selected from the group consisting of extracts for To provide a composition.
  • Dunaliella spp. MP3 Dunaliella sp. MP3, KCTC 12990BP
  • algae algae
  • the culture of the algae its dried products and extracts containing one or more selected from the group consisting of extracts for To provide a composition.
  • the present invention is a food or food additive comprising one or more selected from the group consisting of Dunaliella sp. MP3 ( Dunaliella sp. MP3, KCTC 12990BP) algae, the culture of the algae, dried products thereof and extracts thereof To provide a composition.
  • Dunaliella sp. MP3 Dunaliella sp. MP3, KCTC 12990BP
  • the present invention provides a method for producing pigments comprising culturing Dunaliella sp. MP3 (Dungaliella sp. MP3, KCTC 12990BP) algae.
  • the present invention provides a method of producing a raw material of food or feed comprising culturing Dunaliella sp. MP3 (Dungaliella sp. MP3, KCTC 12990BP) algae.
  • the composition of the present invention can be applied as a raw material of food, health functional foods and pharmaceuticals containing zeaxanthin and lutein pigment.
  • seawater considering the physiological characteristics of Dunariella, a photo-inflammatory microalgae, and the geographical characteristics of Korea, which is a three-sided ocean, seawater can be used as a culture medium, which can be expected to reduce costs and related industrial development.
  • 1A is a micrograph of Dunaliella sp. Algae, wild type (WT) mutant of the present invention, Zea 1 (Zea1), and MP3 mutant strain of the mutant strain of the present invention (magnification 1000x).
  • the bar in the picture represents 10 ⁇ m.
  • 1B is a photograph showing the culture state when wild-type algae, Jia 1, and MP3 mutants were the same number of cells (500 ⁇ 10 4 cells / mL).
  • the upper part shows the culture state in agar containing agar, and the lower part shows the culture state in a liquid culture solution.
  • Figure 3 is a graph showing the cell density (Cell Density) increase pattern of wild type Dunariella genus alga, Jia 1, and MP3 mutant strain of the present invention.
  • the horizontal axis represents the incubation time (day)
  • the vertical axis represents the cell density (x10 4 cells / mL)
  • the number represents the processed light intensity ( ⁇ mol photons / m 2 s), respectively.
  • Figure 4a shows the dry weight per 100 x 10 4 cells (g) of wild-type algae, Jia 1, MP3 mutant strains
  • Figure 4b measures the biomass in dry weight (g) per (L) of wild-type algae, Jia 1, MP3 mutant strains One graph.
  • FIG. 5 is an HPLC analysis graph showing pigment profiles of wild type Dunariella genus algae and Jia 1, MP3 mutant strains (1: neoxanthin, 2: violaxanthin, 3: antheraxanthin, 4: lutein, 5: zeaxanthin, 6: chlorophyll b, 7: chlorophyll a, 8: beta-carotene.
  • FIG. 6a shows the amount of zeaxanthin contained in wild type Dunariella algae, Jia 1, and MP3 mutants, which measured the pigment content of the cells harvested on the 2nd day after incubation at 100 ⁇ mol photons / m 2 s.
  • A shows the content of zeaxanthin per 1 x 10 6 cells ( ⁇ g)
  • B shows the content of zeaxanthin per mg of dry cell weight (mg).
  • Figure 6b shows the pigment content by using HPLC after culturing for 2 days at a luminous intensity of 100 ⁇ mol photons / m 2 s, and measured the pigment content using HPLC, zeaxanthin and lutein contained in wild type Dunariella alga, Jia 1, MP3 mutants, respectively
  • a graph comparing the amount of. shows the content of zeaxanthin and lutein ( ⁇ g) per 1 x 10 6 cells, (B) shows the content of zeaxanthin and lutein (mg) per 1g dry cell weight.
  • Figure 7a is a graph showing the content (wt%) of each pigment in the total amount of pigments produced from wild type Dunariella algae, Jia 1, MP3 mutant strains.
  • 7b is a graph showing the content of zeaxanthin (mg / g DCW) per 1g dry cell weight of Jia 1 and MP3 mutant strains cultured for 14 days.
  • Figure 8a is a diagram showing the PCR results using the primer information and primers of ITS1 -F and ITS2 -R for phylogeny in Dunariella genus.
  • 8B shows a schematic of Dunaliella sp. Strains.
  • the present invention relates to a two Canary in Ella Macular pigment 3 (MP3) (Dunaliella sp. MP3, KCTC 12990BP) algae (algae).
  • MP3 Canary in Ella Macular pigment 3
  • KCTC 12990BP Canary in Ella Macular pigment 3
  • the birds of the genus Dunariella is a single-celled algae with two flagella of isotonic cells, fusiform, and the size is 7 ⁇ 25 ⁇ m in length depending on the type, has a congenital chloroplast. Can grow in environments with high salt concentrations.
  • mutant strains were prepared by general mutagenesis and were isolated using subculture and identified using conventional methods.
  • the mutation treatment can be carried out using common mutagens, and in one embodiment of the invention treated with chemicals to cause the mutation.
  • Dunariella MP3 mutant strain of the present invention has a pigment producing ability, specifically xanthophyll producing ability. More specifically, at least one selected from the group consisting of lutein and zeaxanthin and having chlorophyll b, chlorophyll a and beta-carotene Can produce.
  • the mutant strains were significantly higher in production of zeaxanthin per cell than conventional wild type Dunariella birds, and Dunaliella sp. Zea1, BIOTECHNOLOGY AND BIOENGINEERING , VOL. 81, NO.1 Compared with), the accumulation amount of zeaxanthin per cell is 20 to 30% or more, which can be effectively used as an algae for producing xanthophyll.
  • the MP3 mutant of the present invention is the best accumulation of zeaxanthin per unit cell number and dry weight compared to wild-type and Jia 1 algae of the same genus (Fig. 6b), the mutant strain of the present invention It can be seen that the production ability of xanthophyll, in particular, zizatin has very good bacteriological properties, and it can be effectively used as a source of xanthophyll pigment by using this.
  • the mutant strains of the present invention are viable in dim light and specifically can be cultured under luminous conditions in the range of 10 to 2,000 ⁇ mol photons / m 2 s. Photosynthesis is not possible at complete darkness where the mutant strain is below the weak light condition, and cells may be damaged by light stress at too high light conditions.
  • culturing the mutant strain of the present invention under the above conditions there is an advantage of having an excellent growth rate while increasing the xanthophyll content in the mutant strain.
  • the MP3 mutant strain can grow appropriately within the growth environment (luminosity, temperature, salinity conditions, etc.) of the algae of the genus Dunariella. In addition, it has an excellent ability to accumulate zeaxanthin even at low luminosity (Fig. 6), and can be used industrially effectively as a xanthophyll pigment-producing microorganism due to such excellent xanthophyll production ability, compared to other algae in the density under high brightness. Therefore, it is relatively low, and the pigment
  • the MP3 mutant strain may be cultured in a seawater environment, and specifically, may be cultured in a culture medium including seawater. Variants of the present invention can be cultured at salinity conditions of 0.05M to 5.5M based on NaCl concentration.
  • the culture medium may further include Tris in addition to NaCl.
  • the culture medium may contain a nutritional substance required by the microorganism to be cultured, that is, the culture medium in order to cultivate a specific microorganism, and may be mixed with an additional material for a special purpose.
  • the medium may also be referred to as an incubator or a culture medium, and is a concept that includes all natural, synthetic, or selective media.
  • the Dunaliella mutant strain may be cultured according to a conventional culture method. In one embodiment, in the culture composition of Table 1, it was confirmed that the MP3 mutant strain of the present invention has excellent zeaxanthin production capacity.
  • the pH of the culture medium is not particularly limited as long as the algae in the Dunariella can survive and grow, for example, pH 6 or more, specifically pH 7 to pH 9, and can be optimal at pH 8.0 or more below pH 9.0 It can have a growth rate of.
  • Dunaliella genus 1 (The MP3 variant) sp. Zea1, BIOTECHNOLOGY AND BIOENGINEERING , VOL. 81, NO. It was developed through chemical mutation using 1) as an aid, and identified as Dunaliella sp. By the ITS sequence analysis.
  • ITS1 ITS1 -F: TCCGTAGGTGAACCTGCGG, ITS1 -R: GCTGCGTTCTTCATCGATGC
  • ITS2 ITS1 -F: GCATCGATGAAGAACGCAGC, ITS2 -R: TCCTCCGCTTATTGATATG
  • UPGMA unweighted pair group method with arithmetic mean
  • the selected mutant of the present invention was named Dunaliella sp. Macular pigment 3 (MP3) in Dunariella genus, and the mutant strain was the Korea Research Institute of Bioscience and Biotechnology (KCTC). ) was granted March 16, 2016 and was granted accession number KCTC 12990BP on March 23, 2016.
  • MP3 Macular pigment 3
  • KCTC Korea Research Institute of Bioscience and Biotechnology
  • the MP3 mutant strain of the present invention can accumulate pigments, especially xanthophyll pigments, in a cell at a high content, and may contain a higher content of zeaxanthin, thereby cultivating the algae, such as food, feed, pharmaceuticals, etc. It can be effectively used as a raw material.
  • the present invention relates to a culture of algae of Dunaliella sp. MP3, KCTC 12990BP.
  • culture means a medium in which a specific microorganism is cultured, that is, a culture medium, and the culture includes MP3 of the genus Dunariella.
  • the culture is meant to include both the concentrate of the culture or the dried product of the culture in which the culture medium, such as concentrated and dried after the culture.
  • the culture may include by-products thereof, and the formulation is not limited, and may be, for example, liquid or solid.
  • the medium may contain a nutritional substance required by the microorganism to be cultured, that is, the culture medium in order to cultivate the specific microorganism, and may be mixed with an additional material for a special purpose.
  • the medium may also be referred to as an incubator or a culture medium, and is a concept that includes all natural, synthetic, or selective media.
  • the pH of the medium may be a range capable of growing MP3 algae in Dunariella, for example pH 6 or more, preferably pH 7 to pH 9.
  • the present invention also relates to a composition
  • a composition comprising one or more selected from the group consisting of Dunaliella sp. MP3 of the present invention, Dunaliella sp. MP3, KCTC 12990BP algae, a culture of the algae, dried products thereof and extracts thereof. .
  • composition can be used for the purpose of promoting the health of humans and animals.
  • the variant strain of the present invention has a property of generating and accumulating xanthophyll pigments including zeaxanthin and lutein in the body.
  • the composition may be a pigment composition or a xanthophyll pigment composition.
  • the pigment composition may be that containing 10% by weight to 15% by weight of zeaxanthin relative to the total amount of the pigment contained in the composition.
  • the content of zeaxanthin in total pigments per cell of Dunariella wild type algae, Gia 1 algae and MP3 mutant strains was determined. It was confirmed that the content of zeaxanthin was remarkably high (Fig. 7a).
  • the pigment composition may be used as a raw material for food or feed, and may be used as a preparation for oral administration.
  • the pigment composition or the xanthophyll pigment composition containing the composition or extract may be a composition for oral administration in that it can be supplied orally in food, medicine or feed.
  • compositions for oral administration include powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, and the like, oral formulations formulated using methods known in the art.
  • oral preparations can be obtained by tablets or tablets of sugars by combining the active ingredients with solid excipients and then grinding them and adding suitable auxiliaries and then processing them into granule mixtures.
  • excipients examples include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol and starch, cellulose, including starch, corn starch, wheat starch, rice starch and potato starch, etc. Fillers such as cellulose, gelatin, polyvinylpyrrolidone, and the like, including methyl cellulose, sodium carboxymethylcellulose, hydroxypropylmethyl-cellulose, and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate and the like may optionally be added as a disintegrant.
  • the composition may be added to food or feed to achieve a particular purpose use, in this aspect it may be a food composition, a composition for food additives, a feed composition or a composition for feed additives.
  • the body's health can be maintained or strengthened by the xanthophyll pigments produced by Dunariella genus MP3 algae and accumulated in the cells, in particular zeaxanthin and lutein.
  • the zeaxanthin and lutein may be used as a macular pigment to prevent or improve the degeneration of the macula, and is effective in preventing or improving eye diseases associated with macular degeneration.
  • the zeaxanthin and lutein may be used to enhance or maintain eye health; Macular degeneration prevention or improvement; Preventing or improving deterioration of the eye; Improving or preventing damage to the retina; Anti-aging; Maintaining retinal health; Reduced risk of developing macular degeneration; Or since there is an effect of preventing or improving macular degeneration, the feed or food composition may be used for the prevention or improvement of the symptoms, or for the purpose of the effect.
  • additives is included if all the ingredients are added to the food or feed in addition to the main ingredients, for example, food and pharmaceutical safety is added to the coloring, preservation, etc. in the active active material or processed foods having functionality in the food or feed It may be a food additive as defined by the department.
  • the food may be a health functional food. More specifically, it may be a health functional food for eye health.
  • the food, food additives, feed or feed additives composition does not harm the activity of Dunaliella sp. MP3 (Kalid sp. MP3, KCTC 12990BP) algae of the present invention, the culture of the algae, dried products thereof and extracts thereof. It may further include other active ingredients in a range that does not. In addition, additional components such as a carrier may be further included.
  • Feed composition in the present invention may be prepared in the form of fermented feed, compound feed, pellet form and silage (silage) and the like.
  • the fermented feed includes the MP3 algae of the genus Dunariella of the present invention, the dry cells of the algae, the culture of the algae and extracts thereof, and may be prepared by additionally including various microorganisms or enzymes.
  • the blended feed may be prepared by mixing various kinds of general feed and the Dunariella MP3 algae of the present invention, the dry cells of the algae, the culture of the algae and extracts thereof.
  • Pellet feed may be prepared by formulating the fermented feed or blended feed into a pellet machine.
  • Silage may be prepared by mixing the green feed and Dunariella MP3 algae, the dry cells of the algae, the culture of the algae and / or extracts thereof, but the use of the composition of the present invention is not limited thereto.
  • the composition may be mixed with carriers and flavorings commonly used in the food or pharmaceutical field to form tablets, troches, capsules, elixir, syrups, powders. It may be prepared and administered in the form of a suspension or granules.
  • binders, suspending agents, disintegrating agents, excipients, solubilizers, dispersants, stabilizers, suspending agents and the like can be used.
  • the mode of administration may be oral, parenteral or application, but preferably oral administration.
  • the dosage may be appropriately selected according to the absorbency, inactivation rate and excretion rate of the active ingredient in the body, age, sex, condition of the recipient.
  • the pH of the composition can be easily changed according to the manufacturing conditions, such as a drug, food, etc. in which the composition is used.
  • the composition is 0.001 to 99.99 weight of any one selected from the group consisting of Dunaliella sp. MP3 (KCTC 12990BP) algae, a culture of the algae, a dried product thereof and an extract thereof based on the total weight of the composition. %, Preferably 0.1 to 99% by weight, and the content of the active ingredient can be appropriately adjusted according to the method of use and purpose of use of the composition.
  • Dunaliella sp. MP3 (KCTC 12990BP) algae may be included in the composition itself or in dried form, and the culture of the algae may be included in the composition in concentrated or dried form.
  • the dried means the dried form of the algae or its culture, it may be in the form of powder prepared by lyophilization and the like.
  • the extract refers to an extract obtained by extracting from MP3 algae, its culture solution or dried product thereof, includes an extract using a solvent or the like, and obtained by crushing the MP3 algae of the present invention.
  • the pigment accumulated in the cells of Dunaliella sp. MP3 (KCTC 12990BP) alga of the present invention may be extracted and separated by physical or chemical methods.
  • the extraction process may be performed by a conventional method, for example, by adding an extraction solvent and homogenizing, pulverizing the cells can be extracted the target pigment.
  • the algae crushed material is removed by centrifugation, and the extraction solvent can be removed by distillation under reduced pressure. It may also further comprise a conventional purification process. Since the pigment has a property of being insoluble in water, it can be more easily extracted from the algae of the present invention.
  • Dunariella MP3 mutant of the present invention has excellent production capacity of xanthophyll, in particular zeaxanthin at low brightness, the composition comprising the mutant and its by-products have the effect of improving physical activity, maintaining body function and preventing degradation.
  • the composition of the present invention is for maintaining physical health, specifically, maintaining and preventing deterioration of body function in which the xanthophyll pigment is involved. Or it can be used as a raw material contained in food, medicine or feed for the purpose of improvement.
  • Another object of the present invention is to provide a pigment production method using Dunaliella sp. MP3 (Dungaliella sp. MP3, KCTC 12990BP) algae.
  • Another object of the present invention is to provide a food or feed material production method comprising the step of culturing Dunaliella sp. MP3 (Dungaliella sp. MP3, KCTC 12990BP) algae.
  • Dunaliella spp. MP3 (KCTC 12990BP) algae of the present invention can increase the amount of xanthophyll accumulation in the cultured Dunariella algae, thereby efficiently supplying industrially used raw materials. Can be done.
  • the production method may include culturing Dunaliella spp. MP3 ( Dunaliella sp. MP3, KCTC 12990BP) algae.
  • the production method may further include, after the culturing step, separating the Dunaliella genus (sp.) MP3 algae of the present invention from the culture.
  • the separated algae may be further subjected to processing steps including drying.
  • the production method is a step of extracting pigments from Dunaliella sp. MP3 (Dungaliella sp. MP3, KCTC 12990BP) algae, the culture of the algae, the concentrate of the culture or the dried product of the culture; It may include.
  • the culture may be performed in a medium of salinity condition of 0.05M to 5.5M based on NaCl concentration. It may also be carried out under light conditions, in particular light conditions in the range of 10 to 2,000 ⁇ mol photons / m 2 s.
  • Dunariella MP3 mutant strain of the present invention is excellent in pigment production ability even at low brightness, can increase the content of xanthophyll in the body, it is possible to achieve excellent xanthophyll accumulation without administering high brightness energy, effectively used industrially Can be.
  • the extraction may be performed by a conventional method for extracting a pigment from a microorganism, and examples thereof include an enzyme method, an ultrasonic extraction, a mechanical extraction method, and the like.
  • the production method may further include a concentration step of increasing the content of algae after the cultivation, and drying to further reduce the moisture of the algae after the concentration step.
  • concentration step or the drying step is not necessarily required, and in general, it may be performed using a concentration and drying method or machine commonly used in the field to which the present invention belongs.
  • the production method may be carried out by further comprising a purification step after the extraction step, which may be performed by a conventional purification method in the art.
  • Xanthophyll prepared through the concentration or drying step may be used as a raw material for food, health functional food, cosmetics or drugs.
  • the xanthophyll production method may be performed by employing another method within a range that does not impair the effects of the present invention.
  • the information on the alga and the composition may be mutatis mutandis also for the production method of the present invention.
  • Cells were harvested by centrifugation and then resuspended to give a cell density of 8 ⁇ 10 6 to 10 ⁇ 10 6 cells / mL. Ethylmethane sulphonate) was treated to a final concentration of 0.2 ⁇ M. After incubation for two hours, the EMS used for treatment was washed with liquid medium and centrifuged to harvest only cells. This was again resuspended in a liquid medium and cultured in a dark state for one day, and then transferred to a solid medium added only one thousand to the same composition as the liquid medium of Table 1 and plated. After one week, colonies formed on a solid medium were separated, and a single colony was transferred to the liquid medium of Table 1, and cultured in a liquid. Mutants were selected by characterizing each colony.
  • colonies having a lighter green color than wild-type and algae were selected first, and the relative zeaxanthin content was increased through HPLC analysis of algae of the first selected colonies. High algae were secondary selected.
  • the color of the mutant strain is lighter green and yellow than the wild type Dunariella algae and Jia 1, it was confirmed that the color of the mutant strain is lighter green and yellow in the liquid culture solution at the bottom. .
  • each algae was inoculated at a cell density of 1 ⁇ 10 6 cells / mL in each of the media shown in Table 1, followed by 50 , 100 and 300 ⁇ mol photons / m 2 s at different intensities.
  • the temperature was 25 to 26 ° C., and shaking culture was performed at a speed of 90 rpm.
  • 0.5 mL of the culture solution was taken on each of the first, second, third, fourth, and fifth days after the start of the culture, and the number of cells per milliliter was counted under a microscope using a hemocytometer. The density of cells at different intensities was measured at each time of counting, and the results are shown in FIG. 3.
  • MP3 mutant strains of the cell number was small compared to the wild type and Jia 1 as the culture period elapsed. Comparing the time to reach the same cell number, wild-type algae grew faster by 1 to 1.5 days compared to the MP3 mutant strain, but the final cell density was found to reach a similar level.
  • dry weight is also an important part of industrial use of microalgae, so dry weight was measured to compare their final biomass.
  • samples were inoculated at 100 ⁇ 10 4 cells / mL and harvested on log 2nd day.
  • 1 mL and 2 mL of the incubating suspension were harvested by filtration through a microfilter (filter paper) and dried for 24 hours in a dry oven at 65 ° C. for 1 x 10 6
  • the dry weights of the wild type, Jia 1 and mutant strains of the cells were measured, respectively. Because Dunariella grows in a high-salt liquid medium, the remaining salt remains after harvesting through centrifugation to nourish the dry weight of the cells, so the filter paper was filtered to remove salt.
  • Figure 4a The results are shown in Figure 4a.
  • Colonies generated after mutant treatment were separated into single colonies, continued to incubate, and chromatographic analysis was performed using HPLC for each colony.
  • isolated single colonies were incubated for 5 days at 100 ⁇ mol photons / m 2 s conditions in a liquid medium, specific culture conditions were carried out as in the culture conditions of Example 1.
  • the harvested algae were extracted using 80% acetone, the centrifuged supernatant was filtered again using a nylon filter, and then analyzed by injection into HPLC.
  • the total flow rate of the solvent was 1.2 mL per minute, and uniformly decreased Tris of pH 8.0 to 14%, acetonitrile to 84% to 0%, respectively, from 0 minutes to 15 minutes, Methanol and ethyl acetate started at 2% and increased by 68% and 32%, respectively, by the 15th minute.
  • the solvent ratio is then maintained for 3 minutes (15-18 minutes), then for 1 minute (18-18 minutes), return to the rate at which each solvent was started and then for the remaining 6 minutes. It was kept post-run.
  • the pump was used for Shimadzu LC-20A Prominence, the column was Watera Spherisorb TM S5 (DS1 4.6 x 250 mm, 5 ⁇ m Cartridge Column, USA), the column temperature was maintained at 40 °C.
  • the detector analyzed the data using a photodiode array detector (SPD-M20A, Shimadzu).
  • SPD-M20A photodiode array detector
  • the carotenoid pigments such as zeaxanthin were detected at 445 nm and chlorophyll a at 670 nm. , Denmark) was used as a standard to determine the concentration was determined by using a standard curve for the determination of zeaxanthin and chlorophyll a.
  • FIG. 5 shows an HPLC analysis graph showing a pigment profile in each alga, and a graph quantitatively analyzing the zeaxanthin and lutein content of each algae grown under 100 ⁇ mol photons / m 2 s using a chromatogram. .
  • each bird was counted as 1 ⁇ 10 6 cells or After harvesting only the cells in the culture, the results of comparing the content of zeaxanthin on the basis of 1 g of dry cell weight (DCW: Dry cell weight) are shown in FIG. 6a, and the results of comparing xiaxanthin and lutein content.
  • DCW Dry cell weight
  • the pigment content per DCW is obtained by dividing the pigment content (mg / L) obtained by HPLC by the number of cells per ml (1 ⁇ 10 6 cells / ml) (mg / 1 ⁇ 10 6 cells). a, is calculated in a manner to impart to 1 x 10 6 cells per dry weight (mg / 1 x 10 6 cells ).
  • the MP3 mutant strain of the present invention had a higher zeaxanthin peak (No. 5) compared to wild type algae and algae Jia 1.
  • Jia 1 is a zeaxanthin accumulation mutant, when compared with the MP3 mutant of the present invention, it can be seen that the mutant of the present invention has a higher zeaxanthin production capacity.
  • the MP3 mutant strain accumulated more than 20-fold increased zeaxanthin compared to wild-type, and compared to the original animal, zi-1, increased zeaxanthin by about 40% or more. Accumulating. Therefore, it can be seen that the MP3 mutant strain of the present invention has a significantly higher zeaxanthin production capacity compared with the conventional Dunariella species.
  • the amount of zeaxanthin accumulated in the mutant strains isolated by visual observation and quantitative analysis by HPLC was compared with the amount accumulated in the algae (Jia 1, Zea1).
  • Mutants grown under the same luminosity conditions did not show significant differences in the size of the wild-type Dunariella algae and the algae (Jia 1, Zea1) and the cells themselves (FIG. 1A).
  • MP3 mutant strains were yellower than wild type when inoculated into solid medium and liquid medium at 500 ⁇ 10 4 cells / mL.
  • the mutant strains of the present invention can also be visually confirmed to continuously accumulate zeaxanthin even at relatively low luminous intensities of 100 ⁇ mol photons / m 2 s. These luminous conditions generally do not require a lot of energy. It suggests that there is an advantage to obtain the desired product with high energy efficiency.
  • the mutant strains of the present invention have a high content of accumulated xiaxanthin in algae per unit cell number, even though the growth rate is slower than that of wild-type and zi-algae algae under the same luminous conditions. This means that it can be used as an xanthophyll algae. Therefore, experiments were conducted to determine whether the production capacity of zeaxanthin accumulated in the mutant strains of the present invention can be maintained after culture.
  • the culture was performed by inoculating 1 ⁇ 10 6 cells / mL in a 500 mL volume of the culture medium. Brightness and shaking culture conditions are the same as above. After inoculation, the cells were cultured for 14 days (11-12 days after entering a stationary state), and the cells were harvested to determine the content of zeaxanthin in the total pigment. The results are shown in Figures 7a and 7b.
  • the zeaxanthin production of the mutant strain (MP3) of the present invention is 8.3mg / gDCW 30% more than the 6mg / gDCW of the conventional aid jia 1.
  • Jia 1 which is a wild type and an original animal
  • the content of zeaxanthin in the total pigment was significantly higher, and the yield of zeaxanthin per dry weight was also significantly higher than that of the conventional Aid Jia 1. You can see that it is high. Therefore, it was confirmed that the MP3 mutant strain of the present invention had remarkably excellent zeaxanthin production capacity.
  • microorganisms were identified by analyzing the ITS gene sequence (gene sequencing).
  • the amplified portion was cut out from the agarose gel, and DNA was separated using a gel extraction kit of Cosmo genetech Co., Ltd. Then, Macrogen Co., Ltd. was commissioned to analyze the sequence.
  • the analyzed sequences were compared with the ITS of several strains of Dunaliella, registered at NCBI (http://www.ncbi.nlm.nih.gov/).
  • the ITS sequences of each strain were compared using the clustalW program, and the compared sequences were plotted using the UPGMA algorithm (unweighted pair group method with arithmetic mean) in the MEGA6 program to draw a phylogenetic tree. Comparative analysis.
  • the ITS sequence of Dunariella was compared from the gene bank of NCBI, and the details are as follows (indicated by the gene bank code in parentheses).
  • D.tertiolecta A2 U66956
  • D.tertiolecta UTEX999 AF313434, AF313434
  • D.salina UTEX1644 AF313428, AF313429
  • D.salina CONC006 AF313424, AF313425
  • D.salina CONC007 AF313426, AF313427
  • D. bardawil strain ATCC30861 AF313430, AF313431
  • D. lateralis AF313444, AF313445
  • C. reinhardtii U66954.
  • the selected mutant strain of the present invention (Dunariella genus) Dunaliella sp.), Macular Pigment 3 (MP3) in Dunariella Dunaliella sp. It was named Macular pigment 3 (MP3) and was assigned the accession number KCTC 12990BP from KCTC.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Microbiology (AREA)
  • Wood Science & Technology (AREA)
  • Food Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Botany (AREA)
  • Nutrition Science (AREA)
  • Cell Biology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Animal Husbandry (AREA)
  • Mycology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Physiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medical Informatics (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

La présente invention concerne de nouvelles algues présentant une productivité de pigment améliorée. Un variant selon la présente invention peut être utilisé pour produire des pigments à base de caroténoïde, en particulier, le xanthophylle avec peu d'énergie et permet ainsi de produire efficacement un pigment à un niveau industriel. De plus, la présente invention peut être appliquée en tant que matière première pour des aliments, des aliments fonctionnels de santé et des médicaments contenant des pigments. En outre, lorsque les caractéristiques physiologiques des microalgues euryhalines Dunaliella et les caractéristiques géologiques de la Corée, dont les trois côtés sont en contact avec la mer, sont prises en considération, l'eau de mer peut être utilisée comme milieu de culture, dans l'espoir de réduire le coût et de conduire au développement d'industries apparentées
PCT/KR2017/005685 2016-06-01 2017-05-31 Variant de dunaliella et procédé de production de pigment à l'aide de celui-ci WO2017209510A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN201780034310.7A CN110225968B (zh) 2016-06-01 2017-05-31 杜氏藻突变体和使用其生产色素的方法
US16/306,477 US11179428B2 (en) 2016-06-01 2017-05-31 Dunaliella mutant and method for producing pigment by using same

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2016-0068217 2016-06-01
KR1020160068217A KR101901608B1 (ko) 2016-06-01 2016-06-01 두나리엘라 변이주 및 이를 이용한 색소 생산 방법

Publications (1)

Publication Number Publication Date
WO2017209510A1 true WO2017209510A1 (fr) 2017-12-07

Family

ID=60478801

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2017/005685 WO2017209510A1 (fr) 2016-06-01 2017-05-31 Variant de dunaliella et procédé de production de pigment à l'aide de celui-ci

Country Status (4)

Country Link
US (1) US11179428B2 (fr)
KR (1) KR101901608B1 (fr)
CN (1) CN110225968B (fr)
WO (1) WO2017209510A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102157596B1 (ko) * 2018-10-12 2020-09-18 한양대학교 산학협력단 두나리엘라 배양용 조성물 및 이의 배양방법
KR102107539B1 (ko) 2019-12-18 2020-05-07 송성은 침투이행성과 식이유용성이 증진된 아스타잔틴 및 유용지방산을 포함하는 미세조류 마이크로 분말의 제조방법
KR102508306B1 (ko) * 2020-03-24 2023-03-10 한양대학교 산학협력단 루테인 및 제아잔틴의 함량이 증대된 계란 생산방법, 이로부터 생산된 계란, 및 상기 계란으로부터 난황유 또는 난황 분말을 제조하는 방법
KR20220159073A (ko) * 2021-05-25 2022-12-02 한양대학교 산학협력단 신규 두나리엘라 살리나 및 이의 용도

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060060039A (ko) * 2003-09-17 2006-06-02 니폰 오일 코포레이션 카로티노이드 화합물의 제조방법
KR20100081388A (ko) * 2009-01-06 2010-07-15 한양대학교 산학협력단 신규한 베타-카로틴 생산 균주 및 이를 이용한 베타-카로틴제조방법
CN101597569B (zh) * 2009-05-11 2011-01-05 新奥科技发展有限公司 经甲基磺酸乙酯诱变选育获得的高生长速率杜氏藻

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19838636A1 (de) * 1998-08-26 2000-03-02 Basf Ag Carotinoid-Formulierungen, enthaltend ein Gemisch aus beta-Carotin, Lycopin und Lutein
WO2010045368A2 (fr) * 2008-10-14 2010-04-22 Solazyme, Inc. Compositions alimentaires de la biomasse microalgale
CN101785854B (zh) * 2010-02-11 2012-10-31 石家庄惠仕医药保健品有限公司 叶黄素多维营养保健药物组合物、其制备方法及应用

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20060060039A (ko) * 2003-09-17 2006-06-02 니폰 오일 코포레이션 카로티노이드 화합물의 제조방법
KR20100081388A (ko) * 2009-01-06 2010-07-15 한양대학교 산학협력단 신규한 베타-카로틴 생산 균주 및 이를 이용한 베타-카로틴제조방법
CN101597569B (zh) * 2009-05-11 2011-01-05 新奥科技发展有限公司 经甲基磺酸乙酯诱变选育获得的高生长速率杜氏藻

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
JIN, EONSEON ET AL.: "A Mutant of the Green Alga Dunaliella Salina Constitutively Accumulates Zeaxanthin Under All Growth Conditions", BIOTECHNOLOGY AND BIOENGINEERING, vol. 81, no. 1, 5 January 2003 (2003-01-05), pages 115 - 124, XP002388742 *
MENDOZA, HECTOR ET AL.: "Characterization of Dunaliella Salina Strains by Flow Cytometry: a New Approach to Select Carotenoid Hyperproducing Strains", ELECTRONIC JOURNAL OF BIOTECHNOLOGY, vol. 11, no. 4, 15 October 2008 (2008-10-15), pages 1 - 13, XP055447031 *

Also Published As

Publication number Publication date
CN110225968B (zh) 2024-01-26
US11179428B2 (en) 2021-11-23
CN110225968A (zh) 2019-09-10
KR20170136670A (ko) 2017-12-12
US20190290712A1 (en) 2019-09-26
KR101901608B1 (ko) 2018-09-28

Similar Documents

Publication Publication Date Title
US10973246B2 (en) Chlamydomonas mutants produced using RGEN RNP and method for preparing pigment using the same
WO2017209510A1 (fr) Variant de dunaliella et procédé de production de pigment à l'aide de celui-ci
US20210198680A1 (en) Heterotrophic Production Methods for Microbial Biomass and Bioproducts
WO2022124590A1 (fr) Nouvelle souche schizochytrium sp. et procédé de production d'acides gras polyinsaturés l'utilisant
WO2020045765A1 (fr) Mutant de chlamydomonas et procédé de préparation de celui-ci
US20130309719A1 (en) Heterotrophic microbial production of xanthophyll pigments
US20080293097A1 (en) Biological Production of Zeaxanthin and Carotenoid Biosynthesis Control
KR102445369B1 (ko) 미세조류로부터 기능성 오일을 추출하는 방법 및 기능성 오일을 포함하는 유화액
KR20150071264A (ko) 글루타치온을 고발현하는 신규 균주 및 글루타치온 대량 생산 방법
KR20190100711A (ko) 중성지방 흡수 저해율이 높은 신규 교잡 신품종 버섯 균주 gbn2wp0970 및 그 자실체
US20070190595A1 (en) Process for obtaining zeaxanthin from algae
KR102163257B1 (ko) 비올라잔틴 생산성이 높은 신규 미세조류
WO2020145685A1 (fr) Mutant de chlamydomonas et son utilisation
KR100923105B1 (ko) 아스타잔틴 추출이 용이한 파피아 로도지마 돌연변이주 및그 추출법
US20240101954A1 (en) Novel dunaliella salina and uses thereof
WO2022158814A1 (fr) Nouvelles microalgues présentant une productivité élevée de zéaxanthine et de lutéine
WO2024039163A1 (fr) Nouvelle souche de schizochytrium sp. pour la production de biomasse à haute teneur en protéines, en pigments antioxydants et en acides gras oméga-3, et son utilisation
JP3320417B2 (ja) アスタキサンチン含有動物プランクトン
KR102603612B1 (ko) 아스타잔틴 및 루테인 생산능을 가지는 클라미도모나스속 kiost-2 균주 및 이의 배양방법
KR100249731B1 (ko) 아스타산틴 생산 효모 돌연변이주 및 이의 제조방법
KR0149960B1 (ko) 아스타산틴 함유 효모 제제 및 이의 제조 방법
WO2017183936A1 (fr) Procédé de correction de micro-algues à l'aide de rgen-rnp
EP2157167B1 (fr) Procédé pour obtenir de la lutéine à partir d'algue verte
Barrins et al. Astaxanthin production by Phaffia rhodozyma

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17807003

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17807003

Country of ref document: EP

Kind code of ref document: A1